LU83781A1 - NOVEL 3-AMINO SUBSTITUTED STEROID DERIVATIVES AND THEIR PREPARATION SELSPROCEDES APPLYING TO TAKING MEDICAMENTS AND COMPOSITIONS CONTAINING THEM - Google Patents

Description

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Inventors: descriptive memory '

Cnger DERAEDT, filed in support of a vesperto TORILLï request,

Josette SENZONI. <* e

--------- **"*""' PATENT

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Conv. international; Grand Duchy of Luxembourg Priority cl * a patent application in France New steroid derivatives 3-amino subs- under the number * 30-Î4750. tltue and their salts' Proceae a® Prepa -.- .- --------------------------- ration, application as medicaments and compositions containing them.

• m m · m *. m · _ · M · | M · · · · · M “m _

Société Anonyme known as: ROUSSEL-UCLAF -

The present invention relates to new substituted 3-amino steroid derivatives as well as their salts, the preparation process, the application as medicaments of these new derivatives and the compositions containing them.

5 The subject of the invention is new substituted 3-amino steroid derivatives, as well as their addition salts with mineral or organic acids., Characterized in that they correspond to the general formula I: * 10 in which W represents a hydrogen atom or a hydroxyl radical or, together with X, represents an ethylidene radical, X represents an ethyl radical or a CH3 fH3 .C or CH group or, together with W, an ethylidene- / \ / \ ne radical, the wavy lines mean that the corresponding radical 15 is in one or the other of the configurations oi and ß, represents a hydrogen atom or a methyl radical, R2 represents a hydrogen atom or an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing 2 to 5 carbon atoms and represents a hydrogen atom, a radical alkyl containing 1 to 5 carbon atoms or hydroxyalkyl containing 2 to 5 atones of carbon, an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, or a radical derived from an of-amino acid or from a peptide comprising 2 or 3 ^ -aminic acids, it being understood that: - on the one hand at least one of R3 and does not represent an a-5 hydrogen atom, - d1 on the other hand i) when at: both represents a methyl radical, W and R £ represent a hydrogen atom and X. represents the radical CH- I 3,, in this case ^ R ^ cannot represent a methyl radical, 0 acyl, or a residue derived from an amino carboxylic acid, ii) when at the same time R ^ represents a methyl radical, W represents a hydrogen atom, R2 represents a methyl radical CH-, I 3 t and X represents the radical Cv, in this case, R- cannot represent a methyl or acetyl radical, iii) when at the same time R ^ represents a methyl radical, R2 and W represent a hydrogen atom, X represents a group ch3 i? H whose hydroxyl is of configuration (S) and the

OH

amino radical is in position 3V, in this case, R ^ cannot represent a methyl, acetyl or glycyl radical, iiii) when "- W represents a hydro- CH- \ r I gene, X represents a CH group and the amino radical, is in position 3o <, in this case, the substituents R 1, R 2 and R 3 do not represent at the same time a methyl radical, (iiiii) when at the same time, R 2 and W represent a atom d hydrogen, K represents a methyl radical, X represents a group. · CH, 1 I ^ CH in which the hydroxyl is of configuration (R) and the radical

^ ^ OH

cal amino is in position 3 <x, in this case, R3 does not represent an ethoxycarbonyl, methyl or acetyl radical, iiiiii) when both R ^ and R3 represent a methyl radical and the amino radical is in position 3 < tf, in this case, W and X cannot together represent an ethylidene radical.

3 "*

In the general formula I and in what follows, the term alkyl radical containing from 1 to 5 carbon atoms denotes, for example, a methyl, ethyl, propyl, isopropyl, butyl radical. isobutyl or pentyl; the term hydroxyalkyl radical containing 5 of 2 to 5 carbon atoms denotes, for example, a hy-droxyethyl or hydroxypropyl radical, · the term acyl radical containing 2 to 8 carbon atoms denotes, for example, an acetyl radical, propionyl, n-butyr.yl, isobutyryl, benzoyl or nicotinoyl; the term alkoxycarbonyl radical containing from 2 to 8 carbon-10 carbon atoms denotes, for example, a methoxy, ethoxy or pro-poxycarbonyl radical; the residue derived from an amino carboxylic acid can be chosen from the group consisting of Ala, Val, Ival,

Leu, Ile, Asp, Asn, Glu, Gin, Ser, Thr, Cys, Met, Lys, Arg, Phe, Tyr, Trp, His, and Pro, Nva ', Nie, Hyp, Orn, these acids being below 15 form D or L, as well as by Sar and Gly, all the acids previously named possibly being N-alkylated; when R_ represents

f 'J

a radical derived from a peptide comprising 2 or 3 ^ -aminic acids, these are chosen from the group consisting of the a-cides o <-aminates above.

It will be accepted by convention that the symbols for o <- amino carboxylic acids represent these acids in their D or L configuration (for example, the term Ala means Alanine in D or L form).

Unless otherwise agreed, the nomenclature used in the present application is the IUPAC nomenclature, the rules of which are published in particular in Biochem. J. (1972) 126, 773-780.

The addition salts with mineral or organic acids can be, for example, the salts formed with hydrochloric, hydrobromic, nitric, sulfuric, phosphoric, acetic, formic, propionic, benzoic, maleic, fumaric, succinic, tartaric, citric, oxalic, glyoxylic, aspartic, alkane sulfonic such as methane or ethane sulfonic, arylsulfonic, such as benzene or pa-. sulfonic and arylcarboxylic ratoluene.

35 Among the products which are the subject of the invention, there may be mentioned in particular the derivatives corresponding to formula I above, as well as their addition salts with mineral or organic acids, characterized in that, in said formula I, X represents a CH, I ^

CH group

' OH

4. ’· *

Among these, mention may be made more particularly of the derivatives characterized in that, in said formula I, X represents a CHv group and R2 represents a hydrogen atom.

/ OH

gene or methyl radical, as well as their addition salts with mineral or organic acids and especially: - (2S) 2-amino N - / (20S) -20-hydroxy (5 <tf) -pregnan- 3û <-yl / propana-mide, - le (2S) 2-amino N - / (20S) -20-hydroxy {Soi) -pregnan-3o <-yl / 1H-indol. 3-propanamide, 10 - 2-amino N - / (20S) -20-hydroxy-1.9-nor (5 <x) -pregnan-3o (-yl / acetamide, - 2-amino N- / ( 20S) -20-hydroxy (5 ") - pregnan-3o (-yl / N-methyl acetamide, as well as their salts; the derivatives other than those mentioned above, described in the examples, are also very particularly retained.

A subject of the invention is also a process for the preparation of derivatives, as defined by formula I above, as well as their salts, characterized in that an amine of formula II: CH- is reacted in which the wavy line ^ W, X and I ^ have the meaning already indicated, - either with a halide of formula III:

25 Hal-R'2 III

in which Hal represents a chlorine, bromine or iodine atom, and R '^ represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, to obtain a product of formula I in which R2 represents an atom of hydrogen, 30 R ^ represents an acyl or alkoxycarbonyl radical containing 2 to 8 carbon atoms, and W, X, R ^ and the wavy line have the meaning already indicated, product of formula I which is isolated and salified if desired , - either with an o <-amino acid or a peptide comprising 2 or 3 5 & ^ -amino acids whose amine function is protected by an easily cleavable group in particular by acid hydrolysis, then treated so as to eliminate the protective group, for obtaining a product of formula I in which R2 represents a hydrogen atom, represents a radical derived from an o (-amino or · acid of a peptide comprising 2 or 3 c <-aminic acids, and W, X, R1 and the wavy line have the meaning already indicated, which is isolated and salified if desired é, - either with a halide of alkyl C ^ -C ^ or hydroxyalkyl ren-10 closing from 2 to 5 carbon atoms and whose halogen is a chlorine, bromine or iodine atom, to obtain a product of formula I in which R2 and R ^ represent a hydrogen atom or an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms and W, 15 X, R ^ and the line corrugated have the meaning already indicated, product of formula I which is either isolated and salified if desired, or else, in the case where R ^ represents a hydrogen atom, it is reacted with a halide of formula IV :

Hal-R "3 IV

In which R "3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hal has the meaning already indicated, for obtaining a product of formula I in which R2 represents an alkyl radical C 1 -C 6 or hydroxyalkyl containing from 2 to 5 carbon atoms, R3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms or acyl or acyloxy containing 2 to 8 carbon atoms and W, X, R ^ and the wavy line have the meaning already indicated * 30, which is isolated and salified if desired, or alternatively, in- the case where R3 represents a hydrogen atom, said product of formula I is reacted with an amino acid or a peptide comprising 2 or 3 o-amino acids whose amine function is protected by a group easily cleavable in particular by acid hydrolysis, then treats so eliminating the protective group, to obtain a product of formula I in which R2 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms, R3 represents a radical derived from a tf acid - amino 40 or a peptide comprising 2 or 3 <* - amino acids and W, x, R ^ - * 6 and the wavy line have the meaning already indicated, which is isolated and salified if desired.

Under preferential conditions for implementing the invention, the preparation process described above is carried out as indicated below.

1) The reaction of the amine of formula (II) with the halide of formula (III) is carried out in the presence of an acid-fixing agent, in particular an alkali hydroxide, an alkali carbonate (for example potassium) , an alkaline bicarbonate, an alkaline acetate, an alkaline earth carbonate, a tertiary amine (eg, trialkoylamine or pyridine) or an alkali alcoholate (eg sodium ethylate).

The reaction can be carried out, for example, in inert solvents or suspension media such as dioxane, dime-thylformamide, benzene, toluene or methylene chloride. In the case of acylation, it is also possible to use the free acid, or else a functional derivative other than a halide-nure, such as an acid anhydride.

In the case where free acid is used, it is necessary to carry out the amidation with an activating agent such as a carbodiimide. Other techniques can be used, such as those described for example in "The Peptides" Vol. 1, Academie Press 1979.

2) The reaction of the amine of formula II or of the product of formula 25 in which represents a hydrogen atom and represents an alkyl or hydroxy alkyl radical with the of-amino acid or the peptide whose amino function is protected by an easily cleavable protective group takes place in the presence of a condensing agent. The purpose of the condensing agent in this case is to activate the acid function of the amino acid.

As a condensing agent, a carbodiimide of formula:

A-N = C = N-B

in which A and B represent an alkyl radical containing 35 from 1 to 8 carbon atoms optionally bearing a dialkoylamino radical or represent a cycloalkyl radical.

Mention may be made, for example, of dicyclohexylcarbodiimide or 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide, preferably the latter.

A 2-chloro N-methylpyridinium halide such as iodide can also be used.

It is also possible to use an alkyl chloroformate such as, for example, methyl, ethyl or isobu-tyl chloroformate; it is also possible to use an alkyl pyrophosphite, such as, for example, ethyl pyrophosphite.

As an easily cleavable protecting group, the benzyloxycarbonyl group or the (Z) terbutyloxycarbonyl group (BOC) can be used, as the case may be, for example.

The object of the invention is in particular and a method such as challenge-10 or previously, characterized in that the protective group is the terbutyloxycarbonyl radical.

To remove the above easily cleavable protecting group, an acid such as hydrochloric acid is preferably used as the cleavage agent; the operation is carried out, for example, using an alkanolic solution of hydrochloric acid, or using anhydrous hydrochloric acid by bubbling in ni-methane. Acids such as paratoluene sulfonic acid, formic acid or trifluoroacetic acid can also be used. Hydrogen can also be used in the presence of palladium, for the protective group (Z) for example.

3) The reaction of the amine of formula II with the alkyl or hydroxyalkyl halide is carried out in the presence of the same acid-fixing agents and solvents as in the case of the reaction with the halide of formula III.

The hydroxy function of the hydroxyalkyl radical is advantageously blocked by an easily cleavable protective group, in particular by acid hydrolysis. This protective group is advantageously a tetrahydropyrannyl radical. It can be cleaved under the same conditions as those indicated above for the amino acid protecting group.

4) The reaction of the product of formula I in which represents a hydrogen atom, R2 represents an alkyl radical C ^ -C, - or hydroxyalkyl containing 2 to 5 carbon atoms and W, X, R ^ and the wavy line have the meaning already indicated, with the halide of formula IV is carried out according to the nature of R "^, under the conditions previously described for the reaction of the amine of formula II with the halide of formula III or with alkyl halide.

5) For. the preparation of the products of formula I N-alkylated, 40 it may be advantageous in certain cases to operate in a particular manner:,. 8 - to prepare a product of formula (I) N-monomethylated, one can prepare a carbamate of the amine of the product of formula wire) for example by action of an alkyl haloformate and in particular of ethyl, then reduction of carbamate, for example using aluminum lithium hydride; - to prepare a product of formula (I) N-mono-ethylated, can the corresponding N-acetylated derivative be prepared and then reduced? - to prepare the N-diethyl derivative, simply repeat the operation.

In these three cases, when X represents an 8 -C-CH 4 group, it is necessary to prepare the ketal at 20, for example by the action of ethylene glycol, then, after reduction, to hydrolyze said ketal, for example using a mineral acid.

To prepare a product of formula I N-mono-or di-isopropyl, a product of formula II can be reacted with acetone in the presence of a reducing agent such as sodium cyanoborohydride.

It is obvious that, in the process of the invention, the protection of the amine functions does not concern the cases where these amine functions are alkylated.

The derivatives of formula I with the exception of those for which represents an acyl or acyloxy radical have a basic character. The addition salts of the derivatives of formula I can advantageously be prepared by reacting, in substantially stoichiometric portions, an inorganic or organic acid with said derivative of formula I. The salts can be prepared without isolating the corresponding bases.

The derivatives which are the subject of the present invention have. very interesting pharmacological properties; they are endowed in particular with remarkable immunotherapeutic properties. They are in particular capable of stimulating the immune reactions.

These properties are illustrated later in the experimental part.

These properties justify the use of the substituted 3-amino steroid derivatives of formula (I), as well as their pharmaceutically acceptable salts, as medicaments.

The present application thus also relates to the application, as medicaments, of 3-amino 9 substituted steroid derivatives, as defined by formula I, as well as of their addition salts with pharmaceutically acceptable acids.

Among the drugs which are the subject of the invention,

5 preferably, the drugs characterized in that they consist of the new 3-amino substituted steroid derivatives corresponding to formula I, in which X represents a CEL group

I 3 ment ch as well as by their addition salts with X%

vDH

pharmaceutically acceptable acids.

10 Among these, there are in particular those corresponding to formula I, in which X represents a group fH3 CH, and R2 represents a hydrogen atom or a radical

bH

methyl, as well as their addition salts with pharmaceutically acceptable acids.

15 Among these, the derivatives whose names follow are particularly retained: - 2-amino N - / (20S) -20-hydroxy (5oO -pregnan-3 << - yl / propana-mide, - (2S ) 2-amino N - / (20S) -20-hydroxy (5 "f) -pregnan-3çi-yl / 1H-20 indole 3-propanamide, - 2-amino N - / (20S) -20-hydroxy- 19-nor (5 ".) -Pregnan-3" t * yl / acetamide, - 2-amino N - / (20S) -20-hydroxy (5 <*) -pregnan-3 <* - yl / N-methyl acetamide, as well as their addition salts with pharmaceutically acceptable acids.

The medicaments according to the invention find their use, for example, in the treatment of autoimmune diseases resulting from a deficiency in certain lymphocytes, whether they are diseases of the connective tissue that are not specific for an organ such as, for example, rheumatoid arthritis, systemic lupus erythematosus or whether it is specific organ diseases such as thyroiditis, pemphigus or hemolytic anemia. The medicaments according to the invention can thus be used as an adjuvant treatment for antibiotic therapy and anti-cancer chemotherapy.

The usual dose, which varies according to the derivative used, the subject treated and the condition in question, can be, for example, from 10 mg to 1 g per day, orally in humans, for the derivative of l Example 14 used as an adjuvant for antibiotic therapy.

Finally, the invention relates to pharmaceutical compositions which contain at least one aforementioned derivative or one of its addition salts with pharmaceutically acceptable acids, as active principle.

As medicaments, the derivatives corresponding to formula I and their addition salts with pharmaceutically acceptable acids can be incorporated into pharmaceutical compositions intended for the digestive or parenteral route.

These pharmaceutical compositions can be, for example, solid or liquid and can be presented in the pharmaceutical forms commonly used in human medicine, such as, for example, tablets, simple or coated, capsules, granules, suppositories, injectable preparations, · They are prepared according to the usual methods. The active ingredient (s) can be incorporated therein into excipients usually used in these pharmaceutical compositions, such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous vehicles or no, fatty substances of animal or vegetable origin, paraffinic derivatives, glycols, various wetting agents, dispersants or emulsifiers, preservatives.

The starting materials of formula II, when they are not known, can be prepared as follows:

A product of formula V: in which and the wavy line have the meaning already indicated, is reacted with an excess of ethyl triphenylphosphonium bromide and potassium tert-butylate to obtain a product of formula VI: 11

in which and the wavy line have the meaning already indicated, which is transformed, for example, by passage to to-sylate then action of sodium azide, or directly by reaction with 1 'azidophosphate of diphenyl in the presence · of azodi-5 ethyl triphenylphosphine carboxylate to the azide of formula VII î I

seen

NOT

3 in which and the wavy line have the meaning already indicated and the configuration in 3 is opposite to that of the com-poses V and VI, which are reduced, for example using aluminum hydride-lithium , in order to obtain an amine of formula IIA: H 2 T 2 in which R 1 and the wavy line have the meaning already indicated, whether - either it is isolated, - or it is reduced, for example by hydrogenation in the presence of a catalyst based on Rhodium, platinum or palladium, to obtain a product of formula IIB: Η * »* 20 in which R ^ and the wavy line have the meaning already indicated, - either hydrated for example using diborane to get

CH

nir a product of formula II_ î T 3

yrCU

12 in which and the wavy lines have the meaning already indicated, that either one isolates, or one is subjected to an oxidation for example using chromic acid, to obtain a product of formula II_: p

H2N

in which and the wavy line have the meaning already indicated, - either subject to cis-dihydroxylation, for example using osmic anhydride in the presence of trimethyloxamine,

10 after protection or not of the amine by an easily hydrolyzable group such as a trifluoroacetyl group, to obtain a product of formula II: CH

IT | 3 r4Cbc * “

R

in which R represents the protective group defined above or a hydrogen atom, and R 1 and the wavy line have the meaning already indicated, that either, when R represents a hydrogen atom, one isolates, or, when R represents a protective group, an agent for cleaving said protective group is subjected, for example an alkaline hydrolysis in the case of a trifluoroacetyl group, and then isolates the free amine obtained, or else the it is subjected to an oxidation, for example using chromic acid, to obtain a product of formula 11 ^,: CH ^

R

In which R, R 1 and the wavy line have the meaning already indicated that, either when R represents a hydrogen atom, one isolates, or else, when R represents a protective group, it is subjected to a cleavage agent of said protective group 13, then isolates the free amine obtained, or else it is reduced, for example using sodium borohydride, to obtain a product of formula 11 ^: CH ^ rr '0H ilg l 5 in which R and the wavy lines have the meaning already indicated, that either when R represents a hydrogen atom is isolated - or when R re has a protective group, it is subjected to an agent of cleavage of said protective group, then isolates the free amine 10 obtained.

Examples of preparations of products of formula (II) appear below in the experimental part.

The following examples illustrate the invention without, however, limiting it.

Example 1: (2S) 2-amino N- / (20S) -20-hvdroxy 5oC-preqnan-3oU-yl / propanamide (hydrochloride and base)

Stage A: 2- / l ^ l-dimethyl ethoxy carbonylaml.no/ N - / (20S) -20-hydroxy_ (5 <x) -pregnan-3o <-yl / propanamide

1.92 g of (20S) 3ot-amino 5t * -pregnan-20-ol and 2.27 g of tertbutyloxycarbonyl-L-Alanine (BOC-L-Alanine) are dissolved in 60 cm3 of chloroform and 12 cm3 of pyridine . The solution is stirred at 0 ° C / + 5 ° C under an inert atmosphere and 1.14 g of 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide hydrochloride is added. After 1 hour 15 minutes, 1.15 g of 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide hydrochloride are added again, stirring continued for 50 minutes at 0 ° C / + 5 ° C , brings to dryness, takes up with an aqueous solution of sodium hydrogen carbonate, extracted with ethyl acetate, washed with a saturated aqueous solution of sodium chloride then with a normal aqueous solution of hydrochloric acid and at new with a saturated aqueous solution of sodium chloride, dried, brought to dryness, crystallized from isopropyl ether, drained, washed with isopropyl ether, dried at 60 ° C. under reduced pressure and obtained the expected product. M = 171 ° C.

35 stage B: hydrochloride__ and base_de_j (2S) _ 2-amino N - / (20S) -20- hydroxy 5 <x: -pr egnan- 3p <-yl / _propanamide 14 variant 1: One suspends, under an inert atmosphere, 1.55 g of the product obtained in stage A, in 100 cm3 of nitromé-thane, bubbled hydrochloric acid for 10 minutes, stirred at 20 <'- 25 ° C for 35 minutes, removes excess a -5 hydrochloric acid, wrung, washed with ethyl ether, dried at 60 ° C under reduced pressure, recrystallized from methanol and obtained 990 mg of expected hydrochloride. M 220 ° C. variant 2: 2.7 g of the product obtained in Stage A are dissolved under an inert atmosphere in 10 cm 3 of ethanol, 40 cm 3 of hydrochloric ethalol (3.5N) are added, the mixture is stirred for 8 hours, brought to dryness, resumed at ethyl acetate, ice, drain, wash with ethyl acetate, dry at 60 ° C under reduced pressure, recrystallize from methanol and obtain 2.2 g of the expected hydrochloride.

F "220 ° C.

15 Preparation of the base:

1.74 g of hydrochloride are dissolved in 100 cm3 of tetrahy-drofuran in 30 ° of water, 4 cm3 of 2N sodium hydroxide are added, brought to dryness, taken up in 100 cm3 of chloroform, washed with water, dried, brought to dry, recrystallized from ethyl acetate and obtained 1.27 g of the expected base. M = 224 ° C.

By operating according to a process analogous to that described above, the derivatives described in the table below were prepared.

J (S) t _ R3 _ N ° of „Amino Acid of„ _

‘., - Variant ,, R„ F

1 example start 3

2 1 + Acid BOC-L- O ^ CH-C ^ -C ^ -GOOHCS) 220oC

note Glutamic NE ^ (hydrochloride)

3 1 BOG-L-Serine cAcH-CH-OH (S) 210 ° C

25 (hydrochloride)

4 1 BOC-L-Phenyl- O ^ CH-CHj- ^% (S) 240 ° C

alanine (hydrochloride) 1

1 BOC-L-Trypto- (AcH-CH -n- / V (S) 135 ° C

phan jm 2 (base)

2 I

____H__ 15 N ° of __ ·, Amino Acid of

. i, Variant,. R- F

1 example start 3

6 2 BOC-L-Proline (S) 275 ° C

'1 * (hydrochloride)

1 H (240 ° C

7 CF3C00H BOC-Glycyl O ^ CH -NH-CO-CH -NIL (hydrochloride

Glycine (200 then 252 ° C.

(base

Note for example 2:

The second acid function of L-Glutamic acid is blocked in the form of a benzyl ester; it is released by catalytic 5 · hydrogenation in 11 acetic acid in the presence of palladium, before the cleavage of the BOC.

Example 8: N - / (2QS) -20-hvdroxv 5û ^ -preqnan-3o <-yl / 3-pyridine * carboxamide

Dissolve under an inert atmosphere 960 mg of (20S) 3 <* - amino 10 5 <* - pregnan-20-ol and 813 mg of nicotinic acid in 30 cm3 of chloroform and 6 cm3 of pyridine, add 582 mg of hydrochloride of 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide. After 3 hours 30 minutes, with stirring, 291 mg of 1-ethyl hydrochloride 3— (3-dimethylaminopropyl) carbodiimide is added, stirred for 16 hours, brought to dryness, taken up in 30 cm3 of water, ice, drained, washed with water, dried at 80 ° C under reduced pressure, recrystallized from methanol and obtained 920 mg of the expected product. Mp 258 ° C. / <x / D = + 14.5 ° + 1 ° (c = 1% pyridine).

Example 9: 2-amino hydrochloride N - / (20S) 17tX, 20-dihvdroxv 20 5 <x-preanan 3ot-vl / acetamide

By operating according to a process analogous to that described in Example 1 (variant 2) from BOC-Glycine and (20S) 3 <tf-amino 5 <* - pregnan 11 oc, 20-diol, the product is obtained expected.

M 200 ° C.

25 / <* / D = + 4 ° + 1 ° (0 = 1.5¾ Ethanol 95 °).

The starting (20S) 3 <* - amino 5 <x-pregnan-17 <x, 20-diol can be prepared as follows:: izL (5 ") _ pregn-17 (20) -en 3 ^ -ol On add 59.4 g of triphenylethylphosphonium bromide to 30 16.1 g of potassium tert-butoxide in solution in 160 cm3 of tetrahydrofuran, stir for 30 minutes, add 23.2 g of epiandrosterone, stir for 15 hours, pour into water. * · 16 ice-cold, extracted with ethyl acetate, washed with water, dried, distilled dry, purified by chromatography on silica (eluent: cyclohexane-ethyl acetate 7-3), crystallized in methanol, ice, drain, dry and obtain 23.1 g of the expected product.

5 F = 160 ° C.

Stage 2: (Z) 3ot-azido (5ctf) -pregn; -17 (20) -ene

1.92 g of ethyl azodicarboxylate and 3.02 g of diphenyl azidophosphate are added to a solution of 1.66 g of the above product in 30 cm 3 of benzene and 5 cm 3 of tetrahydro-10 furan, stirred in an ice bath, add in 20 minutes a solution of 2.88 g of triphenylphosphine in 30 cm3 of benzene, stirred again for 40 minutes at 10 ° c, wash with a 2N hydrochloric acid solution, then with water , dried and distilled dry, purified by chromatography on silica (eluent: heptane then 15 heptane-ethyl ether 1-1), and obtained 1.67 g of crystals of the expected product. M = 114 ° C after recrystallization from methanol.

Stage 3: (Z) 5c <-pregn 17 (20) -ene-3 <rt-amine (and hydrochloride)

14.5 g of 20 (Z) 3 <X-azido (5 <*) - pregn.? · Are dissolved in 290 cm 3 of tetrahydrofuran. 17 (20) -ene as obtained above, stirred while heating at 25-27 ° C, added in 1 hour 800 mg of lithium aluminum hydride, stirred for another 1 hour, removed excess hydride with methanol, filter, wash the filtrate with an aqueous solution of Seignette salt then with a saturated aqueous solution of sodium chloride, dry, dry distillate and obtain 13.1 g of crystals of the expected amine. P if 90 ° C.

The base is dissolved in 150 cm3 of ethyl acetate and 30 cm3 of methylene chloride, 27 cm3 of 1.7n hydrochloric ethyl acetate are added, drained, washed with ethyl acetate and • dried under reduced pressure the crystals obtained and obtains 13.2 g of crystals of the hydrochloride of the expected product. Mp 300 ° C. M / -q at 1% in pyridine, at 10¾ of water = + 38.5 ° + 1.5 °.

Stage 4: N - / _ (Z, 5c *) _- pregn 17_ (20) -èn 3 <x-yl / trif luoroacetamide

Is suspended under an inert atmosphere 16.5 g of the chlo-35 hydrate obtained a, u stage..3 above, in 165 cm3 of methylene chloride and 16.5 cm3 of pyridine, cooled to about 5 ° C. , introduced 16.5 cm3 of trifluoroacetic anhydride in 5 minutes, stirred for 15 minutes at room temperature, distilled dry under reduced pressure, added 200 cm3 of water, wrung, washed with 40 water, dried under reduced pressure and obtained 18.1 g of crystals.

'* ”17 F = 204 ° C.

Stage 5: N - / (20S) _ 17o <120 - dihydroxY __ (5 ^) -P £ egnan-3 ^ -yl / _trifluo-roacetamide

18.1 g of the product obtained above are dissolved under an inert atmosphere in 100 cm 3 of methyl ethyl ketone, 9 g of trimethylamine N-oxide dihydrate and a solution of 360 mg of osmium tetroxide in 71 cm 3 of methyl ethyl ketone are added , stirred for 2 hours at reflux, allowed to cool, added 200 cm3 of sodium thiosulfate solution at 10¾ in water, stirred for 10 minutes at room temperature, decanted, washed with water, dried over magnesium sulfate, filter, dry distilled under reduced pressure, purifies the oil obtained by chromatography on silica (eluent: benzene-ethyl acetate 7-3), and obtains 14 g of the expected product; M = 172 ° C then 192 ° C.

Stage 6: (20S) 3 <* - amino 5 <x-pregnan-17 <*, 20 diol, 4 g of the product obtained above are dissolved under an inert atmosphere in 20 cm 3 of methanol, 8 cm 3 of lye solution are added. soda, stirred for 1 hour 30 minutes, added 50 cm3 of water, stirred for 10 minutes, wrung, washed with water, dried under reduced pressure at 40 ° C. and obtained 3 g of sought product. Mp 210 ° C. Example 10: 2-amino hydrochloride N - / (20S) -20-hvdroxv 19-nor (5 <x) -precnan — 3 <* - vl / acetamide

By operating according to a process analogous to that described in Example 1 (variant 1) from BOC-Glycine and (20S) 25 3e <-amino-19-nor (5o <) -pregnan-20-ol, we obtains the expected product. F - 270 ° C with sublimation.

/ <* / D + 39.5 ° + 1.5 ° (c = 1% methanol).

The starting (20S) 3οώ-amino-19-nor (5 & 4) -pregnan-20-ol can be prepared as follows: 30 By operating according to a process identical to that described in Example 9 (stages 1,2,3 ) for the preparation of (Z) 5o <-preg-17 (20) -èn-3o <-amine, we prepared from 19 Nor-epian-drosterone the (Z) 5c <-19 nor-pregn 17 (20) -èn-3 examines.

156 mg of sodium borohydride is suspended in nitrogen in 5 cm3 of tetrahydrofuran, a solution of 0.5 cm3 of boron trifluoride etherate in 2.5 cm3 of tetrahydrofuran is added dropwise at + 5 ° C. hour in an ice bath, add 296 mg of (Z) 5 <x-19 nor-pregn 17 (20) -en-3 <* - amine in solution in 3 cm3 of tetrahydrofuran, stirred for 1 hour 40 30 at room temperature , cooled in an ice bath, added. · - * 18 slowly 2 cm3 of 6N sodium hydroxide, stirred for 5 minutes at room temperature, decanted, extracted the aqueous phase with tetrahydro-furan, washed the organic phase with water, added 4 cm3 of 5N sodium hydroxide, 2 cm3 of hydrogen peroxide at 110 volumes, stirred for 45 minutes, extracted with ethyl acetate, washed with water, dried and distilled to dryness under reduced pressure. The dry extract is taken up in 10 cm3 of methanol with 5 cm3 of hydrochloric acid N, heated for 30 minutes in a water bath at 50 ° C., poured into a saturated solution of sodium bicarbonate, extracted with methylene chloride, washed with 10 water, dry, evaporate under reduced pressure and obtain 257 mg of crystals of the expected product. F 190 ° C.

Example 11: 2-amino hydrochloride N- / 17 <* - hvdroxv 20-oxo 5 <* - preqnan-3o <-vl / acetamide

By operating according to a process analogous to that described in Example 1 (variant 2) from BOC-Glycine and 3 <* - amino 1 ^ -hydroxy 5ctf-pregnan-20-one, the expected product is obtained.

F ’> 300 ° C.

M / o = + 50 ° + 10 (c. = 1% Ethanol 95 °).

The starting 3o <-amino 17oc-hydroxy 5 <x-pregnan-20-one can be prepared as follows:

By perchromic oxidation of the product obtained in stage 5 of Example 9, the N- (17-hydroxy-20-oxo-5 "c-presgnan-3o <-yl) trifluoroacetamide is prepared (M = 178 ° C. then 186 ° C), which is treated as indicated in step 6 of Example 9 to obtain the 3oc-25 amino 17 "c-hydroxy 5o <-pregnan-20-one. M = 216 ° C (after crystallization from water).

Example 12: 2-amino hydrochloride N - / (20R) -20-hvdroxv 5 <* - preanan-Stx'-vl / acetamide

By operating according to a process analogous to that described in Example 1 (variant 1) from BOC-Glycine and (20R) 3 <* - amino 5o <-pregnan-20-ol, the expected product is obtained.

M = 210 ° C then 260 ° C.

/ <x / D = + 22 ° + 1 ° (c = 0.8% pyridine at 10% water).

Example 13: N - / (20S) -20-hydroxy (5cx) -preqnan-3 <* - vl / carbamate 35 d1éthvle

5 g of (20S) 3 "are added dropwise over 10 minutes under an inert atmosphere: - amino 5 <* - pregnan-20-ol dissolved in 500 cm3 of methylene chloride to 15 cm3 of ethyl chloroformate and 45 cm3 of methylene chloride, stirred for 30 40 minutes, added 20 cm3 of sodium hydroxide, stirred again for 30 minutes, decanted, extracted with methylene chloride, washed with water, .. ·. 19 dried, filtered, evaporated to dryness under reduced pressure, taken up in ethyl ether, wrung and obtained 5.7 g of the expected product.

F & 198 ° C.

/ c * / p = + 25 ° + 1.5 ° (c = 1% methylene chloride).

Example 14: 2-amino hydrochloride N - / (20S) -20-hvdroxv 5 * -preqnan-3i * -yl / N-methyl acetamide

By reduction of the product of Example 13, the (20S) 3 <* - methylamino 5t * -pregnan-20-ol (F = 170 ° C) described by Vetter and Coll in Bull was prepared. Soc. (1963) 1324, which was reacted with BOC-Glycine according to a process analogous to that described in Example 1 (variant 2) to obtain the expected product, from which the hydrochloride is prepared. Mp 250 ° C.

Example 15: 2 (S) 2-amino N-methyl N - / (20S) 20-hvd'roxy-5 <Ç-preqnan- 3o (-vl / propanamide) hydrochloride.

* 15 (20S) 3o (-methylamino 5c <-pregnan-20-ol is reacted with BOC-L ^ Alanine according to a process analogous to that described in Example 1. (variant 1) to obtain the expected product , for which the hydrochloride is prepared, mp 270 ° C.

Example 16: 2 (R) 2-amino hydrochloride N-methvl N - / (20S) 20-20 bvdroxv-5t) <- preqnan-3c <-yl / propanamide.

The (20S) 3o6-methylamino-5o <-pregnan-20-ol is reacted with BOC-L-Alanine according to a process analogous to that described in Example 1 (variant 1) to obtain the expected product, of which prepare the hydrochloride. (Sublimation at around 260 ° C).

Example 17: (20S) 3 "4-ethylamino hydrochloride 5oç-preqnan-20-ol.

The (20S) 3x-amino 5 * .- pregnan-20-ol is reacted with ethyl iodide in the presence of sodium carbonate, to obtain the base of the expected product. Mp 129 ° C after recrystallization from ethyl acetate.

Formation of the hydrochloride: the hydrochloride is prepared by adding hydrochloric ethyl acetate and the expected product is obtained. M 270 ° C.

Example 18: 2-amino hydrochloride N-ethvl N - / (20S) 20-hydroxv 35 50 <-preqnan 3 ^ -vl / acetamide.

The base of the product of Example 17 is reacted with BQC-L-Glycine according to a process analogous to that described in Example 1 (variant 1), to obtain the expected product, from which the hydrochloride is prepared. M 230 ° C.

Example 19: (20S) 3x - / (2-hydroxyethyl) amino / 5oc-preqnan-20-ol hydrochloride.

· 20

7 g of (20S) 3ûC-amino 5o £ -pregnan-20-ol and 7 g of sodium carbonate are suspended in 100 cm3 of anhydrous dioxane, 6.6 cm3 of 2- / 2-bromoethoxy / tetrahydropyran are added and brought to reflux for 24 hours under an inert atmosphere. It is cooled to room temperature, diluted with water, extracted with ether, dried, distilled to dryness under reduced pressure, purified by chromatography on silica (eluent: chloroform-methanol-acetic acid 85-5-10) and collected 7.7 g of yellow oil.

Hydrolysis of the protective group.

10 g of the above product are dissolved in 30 cm3 of ethanol and 15 cm3 of 2N hydrochloric acid, stirred at reflux for 1 hour and 30 minutes, cooled, poured onto 100 cm3 of aqueous sodium bicarbonate solution, ice for 15 minutes, spin dry, wash with water, dry at 40 ° C under reduced pressure, dis-15 sout in 350 cm3 of methylene chloride to 10¾ of methanol, - filter, concentrate to about 50 cm3, add 100 cm3 d ethyl acetate, concentrate by half and wring out the precipitate. It is dissolved in 100 cm3 of methanol, filtered, concentrated to around 30 cm3, added 70 cm3 of ethyl acetate, concentrated by half, wrung out, dried at 40 ° C under reduced pressure and obtained 1.65 g of base of the expected product. Mp 215 ° C.

Hydrochloride formation.

1.5 g of the above base are dissolved in 40 cm3 of methanol, a 1.6 N hydrochloric ethyl acetate solution is added to acidic pH, 60 cm3 of ethyl acetate are added, concentrated under reduced pressure, wrung, dried at 40 ° C under reduced pressure and obtained 1.5 g of the expected product. F y. 260 ° C.

Example 20: = <- dimethvlamino N - / (20S) 20-hvdroxv 5o <-preanan-3c <-yl / acetamide.

. The operation is carried out as indicated in stage A of Example 1, using 2.552 g of (20S) 3ç <'- amino 5 * -pregnan-20-ol and 3.350 g of ‘, Ν-dimethyl glycine hydrochloride. 4 g of crude product are obtained, containing the expected derivative and the 0, N-disubstituted derivative. By saponification with sodium hydroxide in methanol, the expected product is obtained. Mp 206 ° C.

/ * / = + 29 ° + 1 ° (C = 1¾ CHC13).

Example 21:

Tablets corresponding to the formula were prepared: -chloride of (2S) 2-amino N - / (20S) -20-hydroxy 5oi-pregnan-4 ° 3 <rf-yl / propanamide ........ ............................ 20 mg; .. .. 21 - excipient q. s. for a tablet finished at ............ 100 mg.

(Detail of excipient; lactose, starch; talc, magnesium stearate).

Example 22: 5 Tablets corresponding to the formula were prepared: - 2-amino N hydrochloride - / (20S) -20-hydroxy 5 <* - pregnan- 3oc-yl / N-methyl acetamide ...... ....................... 15 mg; - Excipient q. s. for a tablet finished at ............ 100 mg.

(Details of excipient: lactose, starch, talc, 10 magnesium stearate).

Pharmacological study --------------------- 1. - Adjuvant of anaphylactic shock Principle: The administration to animals of a product capable of stimulating the activity of immune systems results in increased shock in response to administration of an antigen to which the animal was sensitized.

Male mice weighing 30 to 35 g are sensitized intra-plantar with serum albumin from beef. 8 days 20 later, they receive this antigen intravenously. Under conditions of minimum sensitization, the control animals do not make a fatal shock during this last administration.

The product to be tested is injected intra-plantar, mixed with the antigen: if this product is an adjuvant, it will increase the sensitization and this will result in a fatal shock when administered by the intravenous route.

The dose which causes mortality equal to or greater than 50% of animals is retained as the active dose.

. .. 22 Results: Product of the example Dose per animal in mg 1 0.5 2 ^ 1 4 5 5 5 6 2 7 1 8 5 9 1 10 0.5 13 5 14 0.5 2. - Rosette test to sheep red blood cells Principle: The administration to animals of a product capable of stimulating the activity of the immune systems results in an increase in their capacity to react to the injection of an immunogenic product.

Male rats aged 3 months are sensitized intraperitoneally with sheep erythrocytes (day 0) 7 10 days later (Day 7) their spleen is removed and the splenocytes are brought into contact with sheep erythrocytes: then counts the percentage of leukocytes around which the erythrocytes have formed rosettes.

The product to be studied is administered per os daily 15 from day -1 to day 1.

The dose of product which approximately multiplies by 2 the percentage of rosettes observed in the control animals is considered as an immunostimulating dose.

* '23 Results: Product of the example Dose per animal in mg / kg per os 1 1 5 2 6> 5 8 5 10 5 11 5 14 10 3. - Study of acute toxicity

The lethal LD0 doses of the various compounds have been evaluated, tested after oral administration in mice.

5 DLq is the maximum dose that does not cause death after 8 days.

The results are as follows:

Product of the DLO example in mg / Kg 1,200 2> 400 .3> 400 4> 400 5> 1,000 6> 800 7> 400 9> 600 10> 400 11> 1,000 12> 400 13> 400 ’14> 200

Claims (9)

1. New substituted 3-amino steroid derivatives, as well as their addition salts with mineral or organic acids, characterized in that they correspond to the general formula I: .x 2 \ n R3 - 5 in which W represents a hydrogen atom or a hydroxyl radical or, together with X, represents an ethylidene radical, CH_ CH- L 3 I 3 X represents an ethyl radical or a group £ or CH 0 OH or, together with W, an ethylidene radical, the wavy lines signify that the corresponding radical is in one or the other of the configurations "* and (i, R ^ represents a hydrogen atom or a radical methyl, R2 represents a hydrogen atom or an alkyl radical containing 1 to 5 carbon atoms or hydroxy alkyl containing 2 to 5 carbon atoms and Rg represents a hydrogen atom, an alkyl radical containing 15 mant of 1 with 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, or a radical derived from an o-amino acid or from a peptide comprising 2 or 3 o (amino acids, it being understood that: on the one hand at least one of R2 and R ^ does not represent an a-20 tome d1 hydrogen, - on the other hand: i) when both R ^ represents a methyl radical, W and R2 * represent a hydrogen atom and X represents the radical Ch3 A, in this case, cannot represent an r a- methyl, acyl or a residue derived from an amino carboxylic acid, ii) when at the same time R1 represents a methyl radical, W represents a hydrogen atom, R2 represents a methyl radical and CH-3 X represents the radical ç, in this case, R ^ cannot represent a methyl or acetyl radical, iii) when, at the same time, represents a methyl radical, R2 and W represent a hydrogen atom, X represents a CH- group t ^ CH ^ whose hydroxyl is of configuration (S), and the / OH amino radical is in position 3oi, in this case, R3 cannot represent a methyl, acetyl or glycyl radical, iiii) when - W represents : a drogen hyrr- CH_ I 3 atom, X represents a ch group and the amino radical X \ OH is in position 3o <, in this case, the substituents R-, R_ J · £ "and R ^ do not represent at the same time a methyl radical, iiiii) when, at the same time, R2 and W represent a hydrogen atom, represents a methyl radical, X represents a gro u- ch3 pement J ^ H whose hydroxyl is of configuration (R) and /% OH the amino radical is in position 3 <*, in this case, R3 does not represent an ethoxycarbonyl, methyl or acetyl radical, iiiiii) when at the same time R ^, Rn and R3 represent a methyl radical and the amino radical is in position 3oi, in this case, W and X cannot together represent an ethylidene radical. 2. New 3-amino substituted steroid derivatives according to claim 1, as well as their addition salts with mineral or organic aci-20, characterized in that X represents f3 a CH / Λ OH * 3 group. - New derivatives 3-substituted amino steroids according to claim 2, as well as their addition salts with mineral or organic acids, characterized in that X represents CH- | 3 a group CH and R "represents a hydrogen atom y \ 1 Oh or a methyl radical. 4. Any of the derivatives of formula I whose names follow: - (2S) 2-amino N - / (2Q $) - 20-hydroxy (5oc) -pregnan-3 o <-yl / propana-30 mide, '* * “26 - (2S) 2-amino N - / (20S) -20-hydroxy (5 *) - pregnan-3t * -yl / 1H-indol - 3-propanamide, - 2-araino N - / (20S) -20-hydroxy-19 nor (5oi) -pregnan-3i * -yl / acetamide, 5. 2-amino N - / (20S) -20-hydroxy (5 <*) - pregnan-3 <* - yl / N-methyl acetamide, as well as their addition salts with mineral or organic acids. 5. Process for the preparation of derivatives of formula I, as defined in claim 1, as well as their salts, characterized in that an amine of formula II is reacted: RT fi Ί \, w II in which the wavy line, W, X and have the meaning already indicated, 15. either with a halide of formula III: Hal-R'3 III in which Hal represents a chlorine, bromine or iodine atom, and R'3 represents an acyl or alkoxycarbonyl radical containing 2 to 8 carbon atoms, to obtain a product of formula I in which R2 represents a hydrogen atom, R ^ represents an acyl or alkoxycarbonyl radical containing 2 to 8 atoms of carbon, and W, X, R ^ and the wavy line have the meaning already indicated, product of formula I which is isolated and salified if desired, 25. either with a-: <x-amino acid or a peptide comprising 2 or 3 “i-amino acids” whose amino function is protected by an easily cleavable group, in particular by acid hydrolysis, then tra ite so as to eliminate the protective group, in order to obtain a product of formula I in which R2 represents a hydrogen atom, R3 represents a radical derived from an o-amino acid or from a peptide comprising 2 or 3 Λ -aminic acids, and W, X, R ^ and the wavy line have the meaning already indicated, which is isolated and salified if desired, - either with a C ^ -C ^ alkyl halide or hydroxyalkyl. containing 35 to 2 to 5 carbon atoms and of which the halogen is a chlorine, bromine or iodine atom, to obtain a product of formula · 27 formula 1 / in which R2 and R3 represent a d atom hydrogen or an alkyl radical containing 1 to 5 carbon atoms or hydroxyalkyl containing 2 to 5 carbon atoms, and W, X, R ^ and the wavy line have the meaning already indicated, product of formula I that either it is isolated and salified if desired, or else, in the case where R3 represents a hydrogen atom, it is reacted with a halide of formula IV: Hal-R "3 IV in which R "3 represents an alkyl radical containing from 1 10 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hal has the meaning already indicated, to obtain a product of formula I in which R2 represents an alkyl radical C ^ -C ^ or hydroxyalkyl containing 2 to 5 carbon atoms, R3 represents an alkyl radical containing 1 to 5, r carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, or acyl or acyloxy containing from 2 to 8 carbon atoms and W, X, R ^ and the wavy line have the meaning already indicated, which are isolated and salified if desired, or else -20 core, in the case where R3 represents a hydrogen atom, said product of formula I is reacted with a <tf-amino acid or a peptide comprising 2 or 3 acids examined, the amine function of which is protected by a group easily cleavable in particular by acid hydrolysis, then treated fa to remove the protective group, to obtain a product of formula I in which R2 represents an alkyl radical containing 1 to 5 carbon atoms or hydroxyalkyl containing 2 to 5 carbon atoms, R3 represents a radical derived from an acid at? -amino or a peptide comprising 2 or 3 ot-amino acids and W, X, 30 Rj and the wavy line have the meaning already indicated, which is isolated and salified if desired. 6. Medicaments, characterized in that they consist of the new 3-amino substituted steroid derivatives, as defined by formula I of claim 1, as well as their addition salts with pharmaceutically acceptable acids. 7. Medicines, characterized in that they consist of the new 3-amino substituted steroid derivatives, as defined in one of claims 2 or 3, as well as by their * 28 addition salts with pharmaceutically acids acceptable. 8. Medicines / characterized in that they consist of 3-amino substituted steroid derivatives. , as defined in claim 4, as well as by their addition salts with the 5 pharmaceutically acceptable acids. 9. Pharmaceutical compositions, characterized in that they contain, as active principle, at least one of the medicaments, as defined in any one of claims 6, 7 or 8. r v